Digestion Buffer, DTT extraction
Preparation
Digestion Buffer, DTT extraction
Step 1.
Prepare ingredients
1M Tris (pH 8)
0.5
mL
(10 mM from 1 M stock)
0.5M EDTA STOCK
14.612
mL
(10 mM from 0.034218 M stock)
5M NaCl Stock
1.461
mL
(50 mM from 1.711157 M stock)
10% SDS
2.5
mL
(0.5 % w/v from 10 % w/v stock)
Step 2.
Adjust final volume to 50 mL with distilled water.
Nested recipe inputs
Used in step 1
1M Tris (pH 8)
Step 1.
Add to appropriate container.
Tris Base
12.114
g
Step 2.
Bring final volume to 100 mL with distilled water. Fill media bottle to with ~80% target volume with distilled water.
Step 3.
Add a magnetic stirrer and place on a stirring plate to mix the solution.
Step 4.
Use a pH meter to measure pH. Slowly add concentrated hydrochloric acid (HCl) solution using a Pasteur pipette to reduce the pH to 8.0.
Be careful not to add too much at a time, since the pH will change rapidly.
Step 5.
Bring final volume to 100 mL with distilled water. Once the desired pH has been reached, bring to final volume using distilled water.
Used in step 1
0.5M EDTA STOCK
Step 1.
Prepare ingredients.
EDTA
7.306
g
(0.5 M)
Step 2.
Bring final volume to 50 mL with distilled water.
Used in step 1
5M NaCl Stock
Step 1.
Prepare ingredients.
Sodium Chloride
14.61
g
(5 M)
Step 2.
Bring final volume to 50 mL with distilled water.
Used in step 1
10% SDS
Step 1.
Prepare ingredients.
Sodium Lauryl Sulfate
5
g
Step 2.
Bring final volume to 50 mL with distilled water.